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A simple and practical agglutination assay for the detection of human anti-mouse antibodies

A simple and practical agglutination assay for the detection of human anti-mouse antibodies Background and Objectives: Human anti-mouse antibodies (HAMAs) are relatively common in human serum and may interfere with therapeutic and diagnostic mouse monoclonal antibodies (MoAbs). We developed a simple particle agglutination test (PaGIA) for the detection of HAMAs. Design and Methods: Red-dyed high density particles were coated with monoclonal mouse IgG. These particles were incubated in the reaction chamber of a gel-card together with serum samples obtained from healthy blood donors (n=32), and patients with clinically proven autoimmune thrombocytopenia (AITP; n=26). Positive reactions were defined by a layer of particles on top of the gel or agglutinated particles dispersed throughout the gel matrix. Furthermore, MoAb-coated particles were subjected to flow cytometry and the results were compared with the new HAMA PaGIA. Results: HAMAs were detectable in 33% of serum samples tested (n=58). Results from flow cytometric analysis revealed a high parallel to those obtained by the PaGIA. Interestingly, we observed an increased incidence of HAMAs in AITP patients (42%) compared to healthy blood donors (26%). Interpretation and Conclusion: The new HAMA PaGIA allows a specific and easy, rapid detection of HAMAs and is suitable for large scale testing. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Human Antibodies IOS Press

A simple and practical agglutination assay for the detection of human anti-mouse antibodies

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Publisher
IOS Press
Copyright
Copyright © 2006 by IOS Press, Inc
ISSN
1093-2607
eISSN
1875-869X
Publisher site
See Article on Publisher Site

Abstract

Background and Objectives: Human anti-mouse antibodies (HAMAs) are relatively common in human serum and may interfere with therapeutic and diagnostic mouse monoclonal antibodies (MoAbs). We developed a simple particle agglutination test (PaGIA) for the detection of HAMAs. Design and Methods: Red-dyed high density particles were coated with monoclonal mouse IgG. These particles were incubated in the reaction chamber of a gel-card together with serum samples obtained from healthy blood donors (n=32), and patients with clinically proven autoimmune thrombocytopenia (AITP; n=26). Positive reactions were defined by a layer of particles on top of the gel or agglutinated particles dispersed throughout the gel matrix. Furthermore, MoAb-coated particles were subjected to flow cytometry and the results were compared with the new HAMA PaGIA. Results: HAMAs were detectable in 33% of serum samples tested (n=58). Results from flow cytometric analysis revealed a high parallel to those obtained by the PaGIA. Interestingly, we observed an increased incidence of HAMAs in AITP patients (42%) compared to healthy blood donors (26%). Interpretation and Conclusion: The new HAMA PaGIA allows a specific and easy, rapid detection of HAMAs and is suitable for large scale testing.

Journal

Human AntibodiesIOS Press

Published: Jan 1, 2006

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